蛋白激酶Cζ在人类NSCLC细胞趋化运动中的作用

Role of PKCζ in chemotaxis of human non-small cell lung cancer cells

关键词:  人类非小细胞肺癌  蛋白激酶Cζ  表皮生长因子  趋化

Key words:  human non-small cell lung cancer  PKCζ  EGF  chemotaxis

摘要:

［目的］探讨蛋白激酶Cζ（PKCζ）在表皮生长因子及其受体（EGF/EGFR）诱导的人类非小细胞肺癌（NSCLC）细胞（A549、H1299）趋化运动中所起的作用。［方法］利用趋化实验比较不同化诱因子及不同亚型PKC分子在趋化运动中的作用。［结果］EGF诱导的趋化运动指数比趋化因子CXCL12诱导的高出近3.6倍（P< 0.001）；且用针对不同PKC亚型的抑制剂及PKCζ假底物处理细胞，Chelerythrine chloride对EGF诱导的A549趋化运动阻断具有剂量依赖性（P₁=0.0372, P₂=0.0098, P₃<0.0001）；myr-假底物对A549趋化运动的阻断也具有剂量依赖性（P₁=0.0227, P₂=0.0081, P₃<0.0001），对H1299趋化运动具有阻断作用但无剂量依赖性（P≤0.0066）；其他抑制剂仍可以使EGF诱导出趋化运动。［结论］在NSCLC细胞中，EGF诱导趋化运动的能力强于CXCL12，且PKCζ参与了EGF诱导的趋化运动。

Abstract:

［Objective］To detect the function of PKCζ in the chemotaxis of human non-small cell lung cancer cells induced by EGF or EGFR.［Methods］Chemotactic capacity of A549 induced by EGF was contrasted with which induced by CXCL12 during chemotaxis assay, treatment with different inhibitors of PKC and myr-pseudosubstrate of PKCζ, functions of different isotypes of PKC were observed in the chemotaxis with or without EGF stimulation.［Results］EGF-induced chemotaxis indexes were about 3.6-fold more than those induced by CXCL12(P<0.001). Among different inhibitors, chelerythrine chloride, an inhibitor of all PKC isotypes, impaired chemotaxis of A549 in a dose dependent manner(P₁=0.0372, P₂=0.0098, P₃<0.0001)and so did PKCζ myr-pseudosubstrate (P₁=0.0227, P₂=0.0081, P₃<0.0001), whereas PKCζ myr-pseudosubstrate significantly interrupted chemotaxis of H1299 (P≤0.0066), without a dose dependent manner. Moreover, EGF still elicited chemotaxis in presence of inhibitors of classic and novel PKC.［Conclusions］EGF is a more potent chemoattractant than CXCL12 for human NSCLC cells, and PKCζ is involved in the chemotaxis of human NSCLC cells induced by EGF.